One of the major problems in managing malignant melanoma is predicting the course of disease progression after removal of the primary tumor. Our main hypothesis is that molecular markers can be used for analysis of tumors and blood as surrogates to predict disease progression and outcome, and treatment efficacy well in advance of clinicopathological events. Malignant melanoma progression occurs from accumulation of multiple genetic aberrations that result in progressive genetic instability. In order to translate the detection and prognostic value of these changes efficiently, analysis must be conducted on specimens from well-defined retrospective and prospective clinical trials. Validation of the molecular results will allow better disease stratification and management of malignant melanoma. In the previous grant period we developed multimarker RTPCR assays to identify blood prognostic molecular markers (PMMs) and demonstrated that they could be useful as potential surrogates of subclinical disease progression, and to predict disease recurrence. We will continue to develop new informative PMMs for blood in this proposal. Our hypothesis is that quantitative levels of PMMs which include melanoma-associated antigens (MAAs) and cellular physiologic factors influencing tumor progression/metastasis in melanoma tumors can also be used as surrogates of disease outcome and the efficacy of active-specific immunotherapy (vaccine). Our hypothesis is that DNA markers in primary and metastatic melanoma can also be used as PMMs. We have identified several DNA markers that have shown potential clinicopathological utility. In this renewal we will focus on assessing paired primary/metastatic melanomas to develop different types of PMMs to complement the blood RT-PCR assay as correlates to disease progression and outcome, and treatment failure. The Aims of the project are as follows: 1) Development and assessment of new mRNA PMMs for assessing melanoma patients' tumor and blood; 2) Development and assessment of DNA markers in melanoma tumors as prognostic indicators of disease progression and outcome; and 3) Validation of the clinical and pathological utility of RNA and DNA molecular markers in melanomas from patients entered in the multicenter PMCV Phase III clinical trial. The Aims will take advantage of the JWCI's large melanoma patient referral and comprehensive follow up to provide resources to develop and assess PMMs. The Aims will also take advantage of the randomized multicenter vaccine trial's archived and prospectively collected specimens to validate the PMMs. Identification of molecular markers for melanoma at different stages of disease progression will allow us to develop an "applicable" molecular progression model to aid in management decisions.